Analysis of B-cell epitopes from the allergen Hev b 6.02 revealed by using blocking antibodies.
Hev b 6.02 (hevein), identified as a major allergen from natural rubber latex (NRL), is involved in the latex-fruit syndrome and also acts as a pathogenesis defense-related protein. Its 3D structure has been solved at high resolution, and its linear epitopes have already been reported. However, information about conformational epitopes is still controversial, even though it is relevant for an accurate diagnosis and treatment, as well as for the study of allergen-antibody molecular interactions. We sought to analyze the B-cell epitopes of Hev b 6.02 at a molecular and structural level, using specific recombinant antibodies. We obtained a murine monoclonal antibody (mAb 6E7) and three human single chain fragments (scFvs A6, H8, and G7) anti-Hev b 6.02 that were able to compete for hevein binding with serum IgEs from latex allergic patients. In vitro assays showed that the mAb 6E7 and scFv H8 recognized the area of Hev b 6.02 where the aromatic residues are exposed; while the scFv G7 defined the amino and carboxy-terminal regions that lie close to each other, as a different epitope. The structural modeling of the Hev b 6.02-scFv H8 and Hev b 6.02-scFv G7 complexes revealed the putative regions of two conformational epitopes. In one of these, the aromatic residues, as well as polar side chains are important for the interaction, suggesting that they are part of a dominant conformational epitope also presented on the Hev b 6.02-IgE interactions. Antibodies recognizing this important allergen have potential to be used to diagnose and ultimately treat latex allergy.
Pedraza-Escalona
Martha
M
Instituto de Química, Universidad Nacional Autónoma de México, Circuito Exterior, Cd. Universitaria, Coyoacán, México, D.F. 04510, Mexico.
Becerril-Luján
Baltazar
B
Agundis
Concepción
C
Domínguez-Ramírez
Lenin
L
Pereyra
Ali
A
Riaño-Umbarila
Lidia
L
Rodríguez-Romero
Adela
A
D000485:Allergens / Q000737:chemistry* / Q000276:immunology; D000595:Amino Acid Sequence; D019138:Antibodies, Blocking / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry* / Q000276:immunology; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood; D020315:Latex Hypersensitivity / Q000276:immunology*; D008969:Molecular Sequence Data; D037121:Plant Lectins / Q000737:chemistry* / Q000276:immunology; D011487:Protein Conformation; D016415:Sequence Alignment
D000485:Allergens; D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D023181:Antimicrobial Cationic Peptides; D018985:Epitopes, B-Lymphocyte; D037121:Plant Lectins; C045875:hevein; D007073:Immunoglobulin E